| Objective:Adenosine triphosphate(ATP) can promote the repairation of spinal cord injury (SCI). To investigate the effect of acute spinal cord injury repaired by ATP combing with bone marrow mesenchymal stem cells (BMSCs) transplantation, and to evaluate the synergistic action of ATP and BMSCs in the repairation of SCI and the feasibility of the combined transplantation in the treatment of SCI.Methods BMSCs were isolated from a male SD rat (weighing 120g) using the marrow of the tibia and the femur, the third generation BMSCs were proliferated and labeled with BrdU, then made into BMSCs suspension of 5.0×107/mL.48 adult female SD rats(weighing 240-260g) were used to make acute SCI models at T11～13 level according to the improved Allen's method, were randomly divided into 4 groups(groups A, B, C and D, n=12). the group of ATP-BMSCs transplantation(group A):Inject ATP(40mg/kg) and BMSCs(6μl) to the sentral point and the other two points which were lmm from the each side of head and tail of the injured spinal cord after blending the BMSCs suspension, the cells amount was about 3.0×105; BMSCs transplantation(group B), ATP treatment(group C) and PBS control(group D):Transplant the BMSCs suspension(6μl), ATP(40mg/kg) and PBS(40 mg/kg) to the points as the metods of group A. Observated the general conditions of the rats after opration; The nerve function of low extremities was evaluated using the promoted Tarlov scale and the Rivilin inclined plane test at 1, 3,7,14,21 and 28 days after transplantation; All the rats were execute 28 days after operation, the reparative effect of SCI was observated using histological and immunohistochemical staining.Results The one of group A, two of group B and C and there of group D died as the result of infection and anorexic, the others survived to the end of the experiment. The animals of every group appeared paralysis symptom in low extremities after operation and which began to recover since 1-2 weeks postoperatively, the group A was the best, group B and C were better, group D is the worst. The promoted Tarlov scale and the Rivilin inclined plane test shew that the comparation among each group 1 and 3 days postoperatively existed no statistical significance(P>0.05); the comparation between group A,B,C and D 7,14 and 21 days postoperatively existed statistical significance(P<0.05).28 days after opration:HE staining demonstrated that the injured region in group A restored good, without obvious scar tissue and cavity, and there existed clear stem cell differentiation characters; there was small amounts of scar tissue and cavity filled in the injury site of group B and C; there was great deal of scar tissue filled in the injury site in group D, in which there were numerous inflammatory cells and fibroblasts infiltration and bigger cavity. Immunohistochemical staining showed that the expression of NF and GFAP in group A was higher than group B,C and D, group B and C was higher than group D, the comparations existed statistical significance(P<0.05), there was no statistical significance between group B and C (P>0.05).Conclusions There exists a protective function in ATP after SCI; Combination of ATP with BMSCs can synergistically promote the reparation of acute SCI. |
PDF Full Text Request